Cancer treatment apparatus

ABSTRACT

A cancer treatment apparatus including a magnetic field generator that generates a magnetic field of 100 kHz to 300 kHz to be applied to affected tissues.

TECHNICAL FIELD

The present invention relates to a cancer treatment apparatus forsuppressing proliferation of cancer cells by applying an alternatingmagnetic field to affected tissues.

BACKGROUND ART

Conventionally, there have been proposed apparatuses for applying anelectric field to the affected tissues as cancer treatment apparatusesfor highly malignant cancers. PTL 1 discloses an apparatus whichselectively destroys or inhibits the growth of rapidly dividing cellslocated within a target region of a patient and which includes: an ACvoltage source; and a plurality of insulated electrodes that areoperatively connected to the AC voltage source, wherein each of theelectrodes has a surface configured for placing against the patient'sbody; wherein the AC voltage source and the electrodes are configured sothat, when the electrodes are placed against the patient's body, a firstAC electric field having a first frequency and a second AC field havinga second frequency are imposed sequentially in terms of time in thetarget region of the patient, wherein the first frequency and the secondfrequency are different, wherein the first and second electric fieldshave frequency characteristics that correspond to a vulnerability of therapidly dividing cells, wherein the first and second electric fields arestrong enough to damage, during the late anaphase or telophase stages ofcell division, a significant portion of the rapidly dividing cells whoselong axis is generally aligned with the lines of force of the electricfields, and wherein the first and second electric fields leavenon-dividing cells located within the target region substantiallyunchanged.

Moreover, PTL 2 proposes an in-vitro method of selectively destroying orinhibiting the growth of parasites located within a target region(1620), including the steps of: capacitively coupling an AC electricfield into the target region (1620); and repeating the coupling stepuntil a therapeutically significant portion of the parasites die withinthe target region (1620), wherein the frequency of the electric field isbetween 10 MHz and 20 MHz, wherein the strength of the electric field inat least a portion of the target region (1620) is between 0.5 V/cm and10 V/cm, wherein the electric field damages or disrupts a significantportion of the parasites positioned within the target region (1620), andwherein the electric field leaves non-dividing cells located within thetarget region (1620) substantially unharmed.

However, with the cancer treatment apparatus which applies the electricfield to the affected part, it is necessary to shave the affected partin order to mount electrode section of the apparatus. Additionally,duration to wear the electrode sections is long and it is sometimesnecessary to wear the electrode sections for 18 hours or longer.Furthermore, if a patient moves during the time when they wear theapparatus, they are forced to carry a heavy power source. In light ofthe current use situation of the cancer treatment apparatus, there is ademand for a cancer treatment apparatus which not only has thetherapeutic effects, but also is suited for a remission treatment withless burden on patients.

There is another cancer treatment apparatus proposed by PTL 3, that is,a ceramic heating element for a thermotherapy characterized in that theceramic heating element is ferromagnetic ferrite particles covered witha bioactive inorganic layer and has a very good affinity for surroundingtissues when embedded in a body and generates heat via magneticinduction highly efficiently within an alternating magnetic field.Furthermore, PTL 4 discloses a cancer treatment apparatus characterizedin that it includes: a pair of magnetic poles for generating analternating magnetic field, wherein the magnetic poles are set atpositions opposite each other to sandwich an affected part in a mannersuch that a clearance between them can be freely adjusted; a magneticfield application unit for applying the alternating magnetic field tothe pair of magnetic poles; and a magnetic field control module forcontrolling the alternating magnetic field.

The inventions described in PTL 3 and PTL 4 are embedded in the body fora long period of time and thereby intended for the cancer treatment bymeans of their hyperthermia effects. However, specific cancer cellcytostatic effects are not disclosed. Therefore, it is desired that thecancer treatment by application of the magnetic field be further studiedand a cancer treatment apparatus which exhibits revolutionary effects onhighly malignant cancer cells be established based on achievements ofsuch study.

CITATION LIST Patent Literature

-   PTL 1: Japanese Patent No. 4750784-   PTL 2: Japanese Patent No. 5485153-   PTL 3: Japanese Patent Application Laid-Open Publication No.    H02-88059-   PTL 4: Japanese Patent Application Laid-Open Publication No.    H03-158176

SUMMARY OF THE INVENTION Problems to be Solved by the Invention

It is an object of the present invention to provide a low-cost cancertreatment apparatus suited for a remission treatment.

Means to Solve the Problems

A cancer treatment apparatus according to a first embodiment of thepresent invention includes a magnetic field generator that generates amagnetic field of 100 kHz to 300 kHz to be applied to affected tissues.The present invention is the cancer treatment apparatus which appliesthe magnetic field of 100 kHz to 300 kHz by placing the magnetic fieldgenerator opposite the affected tissues. The present invention cansuppress the proliferation of cancer cells by applying the magneticfield from the magnetic field generator to the affected tissues. Themagnetic field generator according to the present invention can applythe magnetic field, which is required to obtain a specified operationaladvantage of the present invention, to an affected part even in a statenot in contact with the affected part. Accordingly, the presentinvention should preferably be used by placing the magnetic fieldgenerator opposite the affected tissues in a non-contact state.Incidentally, the temperature of the magnetic field generator upon thegeneration of the magnetic field is approximately from 30° C. to 39° C.,inclusive, so that even if the magnetic field generator contacts theaffected tissues, the affected tissues can be treated safely. Therefore,the present invention can be also used in the state where the affectedtissues and the magnetic field generator are placed opposite each otherand in contact with each other.

The present invention contains the cancer treatment apparatus stated inclaim 1, which further includes: a control module that controls anelectric current supplied to the magnetic field generator; and a powersource that supplies an alternating current to the magnetic fieldgenerator on the basis of output from the control module, wherein thecontrol module includes: a cancer type input unit that accepts input ofa cancer type of the affected tissues; a storage unit that stores afrequency corresponding to the cancer type; and a controller that refersto the storage unit and sets the frequency corresponding to the cancertype, which is input to the cancer type input unit, to the power source.

The present invention contains the cancer treatment apparatus whichfurther stores application duration corresponding to the cancer type;and the controller refers to the storage unit and outputs information ofthe application duration corresponding to the cancer type, which isinput to the cancer type input unit, to the power source. Accordingly,the present invention can simplify the input of the application durationby an operator and contribute to the reduction of a patient's burden.The present invention contains the cancer treatment apparatus whichincludes a power switch that starts supplying the alternating currentfrom the power source to the magnetic field generator. The presentinvention contains the cancer treatment apparatus which controls atemperature of the affected tissues to make it lower than a cancer cellkilling temperature. Accordingly, the proliferation of the cancer cellscan be suppressed at a temperature lower than the cancer cell killingtemperature. The present invention contains the cancer treatmentapparatus regarding which the affected tissues are human affectedtissues.

The present invention contains the cancer treatment apparatus whichincludes a cancer type input unit capable of selecting and inputting onecancer type from a group consisting of a glioblastoma, a malignantmelanoma, a tongue cancer, a breast cancer, a malignant mesothelioma, apancreatic cancer, and a human alveolar basal epithelial adenocarcinoma.Regarding cancer treatments, preferred alternating magnetic fieldconditions to obtain therapeutic effects vary depending on the cancertype. The present invention can intuitively input the frequency andapplication duration which are suited for the treatment target cancertype and contribute to the reduction of the operator's burden.

According to the present invention, the application duration shouldpreferably be configured as a value within a range of 30 minutes to 180minutes, inclusive.

Advantageous Effects of the Invention

The present invention can apply a specific alternating magnetic field,which is suited for suppressing the proliferation of the cancer cells,to the affected tissues. Regarding the present invention, the magneticfield generator which applies the magnetic field to the affected tissuescan be a non-contact type. The application duration is short andadditionally the apparatus is lightweight. Therefore, the presentinvention is suited for the remission treatment.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 is a conceptual diagram of an example of a cancer treatmentapparatus according to the present invention;

FIG. 2 is an example of a flowchart of the cancer treatment apparatusaccording to the present invention;

FIG. 3 is an example of a cancer type input unit according to thepresent invention;

FIG. 4 illustrates shape examples of a coil(s) used for a magnetic fieldgenerator according to the present invention;

FIG. 5 is an application example of the present invention;

FIG. 6 is a simulation result of magnetic flux density regarding amagnetic field generator according to a example of the presentinvention;

FIG. 7 shows observation results of normal cells to which the example ofthe present invention is applied;

FIG. 8 shows observation results of a cancer cell cytostatic effectaccording to the example of the present invention;

FIG. 9 shows observation results of the cancer cell cytostatic effectaccording to the example of the present invention;

FIG. 10 shows observation results of the cancer cell cytostatic effectaccording to the example of the present invention;

FIG. 11 shows observation results of the cancer cell cytostatic effectaccording to the example of the present invention;

FIG. 12 shows observation results of cancer cells to which the exampleof the present invention is applied;

FIG. 13 shows observation results of the cancer cells to which theexample of the present invention is applied;

FIG. 14 shows observation results of a tumor model to which the exampleof the present invention is applied;

FIG. 15 shows observation results of changes in a tumor volume accordingto the example of the present invention;

FIG. 16 shows observation results of changes in the tumor volumeaccording to the example of the present invention;

FIG. 17 is an example of the cancer type input unit according to thepresent invention;

FIG. 18 shows observation results of tumor models according to theworking example of the present invention;

FIG. 19 shows observation results of changes in the tumor volumeaccording to the example of the present invention;

FIG. 20 shows observation results of the cancer cell cytostatic effectaccording to the example of the present invention;

FIG. 21 shows observation results of changes in the tumor volumeaccording to the example of the present invention;

FIG. 22 shows observation results of changes in the tumor volumeaccording to the working example of the present invention;

FIG. 23 shows observation results of an overall survival rate accordingto the example of the present invention;

FIG. 24 shows observation results of side effects according to theexample of the present invention;

FIG. 25 shows observation results of the side effects according to theexample of the present invention;

FIG. 26 shows observation results of the side effects according to theexample of the present invention;

FIG. 27 is a conceptual diagram of another example of the cancertreatment apparatus according to the present invention;

FIG. 28 is a conceptual diagram of further another example of the cancertreatment apparatus according to the present invention; and

FIG. 29 shows observation results of reactive oxygen according to theexample of the present invention.

DESCRIPTION OF EMBODIMENTS

The present invention treats cancer without a use of any heat generationmedium. The heat generation medium referred to here is, for example, aheating element that exhibits porcelain induction heat generation by amagnetic field. An embodiment of the present invention will be explainedwith reference to FIG. 1 and FIG. 2. FIG. 1 is a conceptual diagram ofan example of a cancer treatment apparatus according to the presentinvention. FIG. 2 is a flowchart of the cancer treatment apparatusaccording to the present invention. Referring to FIG. 1(a), 100represents a cancer treatment apparatus, 200 represents a magnetic fieldgenerator, 300 represents a power source, and 400 represents a controlmodule. The magnetic field generator 200 includes a cooling mechanism,such as cooling fans, which is not illustrated in the drawing andmaintains an appropriate temperature of the magnetic field generator200.

FIG. 1(b) is a conceptual diagram for explaining the control module 400illustrated in FIG. 1(a). 401 represents a cancer type input unit, 402represents a power switch, 403 represents a first input unit, 404represents a second input unit, 501 represents a controller, 502represents a memory, 503 represents a first output unit, 504 representsa timer, and 505 represents a second output unit. The control module 400includes a CPU that is a central processing unit which is notillustrated in the drawing, a ROM which is a read-only storage area, anda RAM which is readable and writable; and the CPU implements theoperation described later by expanding programs stored in the ROM ontothe RAM and executing them. However, the control module 400 mayimplement the operation described later via a hardware circuit.

FIG. 2 explains an example of applying a magnetic field by setting acancer type parameter which is a combination of a frequency parameterand an application duration parameter. The frequency parameter and theapplication duration parameter which are stored in the memory 502 can beevoked and desired frequency and application duration can be set byactivating the present invention, selecting a disease name and a cancertype, and inputting them to the cancer type input unit 401. FIG. 3illustrates an example of the cancer type input unit. Referring to FIG.3, a disease name column corresponds to the cancer type input unit 401.The cancer type input unit 401 should preferably display the diseasename from the viewpoint of convenience for the operator; however, theinvention is not limited to this example and, for example, a cancer typemay be displayed.

Regarding numerical values indicated in columns of the frequency and theapplication duration in FIG. 3, preferred values corresponding to thedisease name and the cancer type are associated with such numeralvalues, so that they do not have to be displayed in the cancer typeinput unit. However, the present invention does not exclude anembodiment to display these values in visually recognizable positions.Incidentally, a preferred mechanism for inputting data to the cancertype input unit should include devices such as a keyboard and a mouse, atouch panel, and so on.

Furthermore, when it is designed so that a plurality of applicationmodes can be set for the same disease name, it is also preferable that amode name be assigned to the disease name as illustrated in FIG. 3. Thefollowing explanation will be provided by taking an embodiment to set“Glioblastoma Mode 2” as an example.

When “Glioblastoma Mode 2” is input to the cancer type input unit 401(S610), the cancer type parameter which is a combination of thefrequency of 227 kHz and the application duration of 30 minutes per oneapplication is evoked from the memory 502 via the first input unit 403and the frequency N and the application duration T_(d) are setrespectively upon application of the magnetic field (S620). Afterplacing the affected part opposite the magnetic field generator 200 andswitching the power switch 402 to ON, the power source 300 is notifiedof supply of an alternating current at the frequency of 227 kHz via thefirst output unit 503 and the second input unit 404 from the powersource 300 to the magnetic field generator 200 and the supply of theelectric current is started (S630). Consequently, the magnetic fieldgenerator 200 starts applying the alternating magnetic field to theaffected tissues. Incidentally, at the same time as the start of theelectric current supply, the timer 504 starts measuring elapsed timeT_(v) from the start of the electric current supply (S640). The electriccurrent continues to be supplied until the elapsed time T_(v) becomesequal to the application duration T_(d) (30 minutes in this embodiment),that is, while T_(v)≠T_(d) (S660). After the elapse of 30 minutes fromthe start of the electric current supply, that is, when the elapsed timeT_(v) becomes equal to the application duration T_(d) (T_(v)=T_(d))(S670), the power source 300 is notified of the completion of theapplication duration via the second output unit 505, the electriccurrent supply to the magnetic field generator 200 is stopped, and theapplication of the magnetic field to the affected part is terminated(S680). Regarding stoppage of the electric current supply, a mechanismfor forcibly stopping the supply may be provided within the apparatus,or a notification means such as an alarm may be provided and the supplymay be stopped manually.

The frequency parameter which is stored in the memory according to thepresent invention should preferably be a value within the range of 100kHz to 300 kHz, inclusive. When the frequency parameter is less than 100kHz, any significant cancer cell cytostatic effect will not be obtainedeven if the magnetic field is applied. When the frequency parameterexceeds 300 kHz, there is a possibility that the effect might attenuate.Also, as compared to the case where the frequency parameter is equal toor less than 300 kHz, there is a possibility that the temperature of themagnetic field generator might increase. The frequency at which thecancer cell cytostatic effect may tend to appear easily varies dependingon the cancer type of a magnetic field application target. Regardingvalues stored in the memory as the frequency parameter for aglioblastoma, a malignant melanoma, a tongue cancer, a breast cancer,and a malignant mesothelioma, it is possible to store values which aredispersed at regular intervals, for example, every 50 kHz, that is, 100kHz, 150 kHz, 200 kHz, 250 kHz, and 300 kHz. Alternatively, specificvalues such as 196 kHz, 227 kHz, 280 kHz, and 389 kHz may be stored orthe respective parameters may be dispersed at irregular intervals andstored with reference to a specified frequency for each cancer type. Forexample, it is an embodiment where a relatively small interval(s) is setaround a most preferred value and a relatively large interval(s) is setaround a maximum value and a minimum value.

The application duration parameter stored in the memory according to thepresent invention should preferably be a value within the range of 30minutes to 180 minutes, inclusive. When the application duration is lessthan 30 minutes, any significant cancer cell cytostatic effect will notbe obtained even if the magnetic field is applied. If the presentinvention is used for 30 minutes or longer, the significant cancer cellcytostatic effect will be obtained even if the application duration isequal to or less than 180 minutes. Therefore, when using the presentinvention, it is unnecessary to apply the magnetic field to the affectedpart for a long period of time and the patient's burden can be therebyreduced. Additionally, since operating time of the present invention perpatient is short, a plurality of patients can take turns using oneapparatus of the present invention in one day. Furthermore, since thepresent invention can be used in the state of placing the affectedtissues not in contact with the magnetic field generator, hygienicmaintenance required when the user is switched is easy. As a result, thepresent invention contributes to cost reduction.

The application duration which brings about the cancer cell cytostaticeffect in a preferable manner varies depending on the cancer type andthe frequency. When the application duration is stored in the memory asthe application duration parameter for the glioblastoma, malignantmelanoma, tongue cancer, breast cancer, and malignant mesothelioma, itis possible to store, as an example, 30 minutes, 45 minutes, 60 minutes,120 minutes, and 180 minutes. The frequency and the application durationmay be set individually. In that case, a first cancer type input unitfor setting the frequency and a second cancer type input unit forsetting the application duration may be provided. In that case, as aresult of input to the first and second cancer type input units, thefrequency parameter and the application duration parameter are evokedindividually from the memory and the specified frequency and applicationduration are set respectively. From the viewpoint of convenience of thepresent invention, it is preferable as explained with reference to FIG.3 that a combination of the frequency and the application duration beset as the cancer type parameter.

Furthermore, it is also preferable that according to the presentinvention, the number of times of applications be set according to thesize of the relevant tumor, its progression, and the condition of thepatient such as their physical condition. Therefore, it is alsopreferable that the present invention be configured to be capable ofstoring a parameter of the number of times of applications in the memoryand setting the number of times of applications by making an input tothe cancer type input unit. Furthermore, if the cancer type parameter iscombined with the parameter of the number of times of applications, thefrequency parameter, and the application duration parameter, not onlythe frequency and the application duration, but also the number of timesof applications can be set by one input. Accordingly, the presentinvention can prevent errors in a magnetic field application scheduleand further reduce the user's burden of operation.

The number of times of applications according to the present inventionshould preferably be once a day or may be one day per week or alsopreferably be consecutive five days or more per week. For example, whenthe number of times of applications is set as consecutive five days, themagnetic field can be applied according to the present invention bysetting the application start day as the 1st day, applying the magneticfield once every day from the 1st day to the 5th day, and not applyingthe magnetic field on the 6th day and the 7th day. Since the applicationduration per one application according to the present invention isshort, it hardly becomes the user's burden even if the number of timesof applications increases. Furthermore, the definition of the number oftimes of applications is not limited to the number of times per week.Specifically speaking, the number of times of applications may be setfor a cycle such as per five days or per two weeks.

It is preferable that a coil be used as the magnetic field generatoraccording to the present invention. Examples of the shape of the coilcan include a helical type and a disc type. FIG. 4 illustrates examplesof the coil shape used for the magnetic field generator according to thepresent invention. Referring to FIG. 4, FIG. 4(a) is a helical-type coilobtained by molding a metal wire in a helical shape; FIG. 4(b) is adisc-type coil obtained by molding the metal wire in a spiral shape;FIG. 4(c) is a double-disc-type coil formed by placing two disc-typecoils illustrated in FIG. 4(b) side by side. When the coil of any one ofthe shapes is used, the present invention can suppress the proliferationof the cancer cells by applying the magnetic field without placing themagnetic field generator and the affected tissues in contact with eachother.

The magnetic field generator according to the present invention may beconfigured so that the coil of a single shape is secured; however, it ispreferable that coils of a plurality of shapes be made replaceable.Accordingly, the magnetic field generator can be formed by selecting andattaching the coil of a preferred shape to the site of the affectedpart, depending on whether the affected part is located at any one of ahead, four limbs, a torso, and other sites. As a result, the magneticfield can be applied more efficiently to the affected part. The presentinvention can be used as it is placed opposite the affected tissues andwithout contacting the affected tissues. Additionally, as the magneticfield generator includes a replaceable coil, its versatility can befurther enhanced. Also, the coil to be secured to the magnetic fieldgenerator may be a combination of a plurality of coils with differentproperties. FIG. 5 illustrates an application example of the presentinvention. FIG. 5 illustrates an embodiment in which the magnetic fieldgenerator including the helical-type coil is placed opposite anindividual (mouse) having an affected part in its torso and thealternating magnetic field is applied to the individual.

Incidentally, the control module 400 may not include the power switch402 or the timer 504. In this case, the configuration of the controlmodule 400 is as illustrated in FIG. 27 and the control module 400 maynot include the second input unit 404 or the second output unit 505. Inthis case, the control module 400 can implement the same operation asthe case where the power switch 402 and the timer 504 are included, bycontrolling the electric power supplied to the power source 300 fromoutside the cancer treatment apparatus 100. Furthermore, the controlmodule 400 may not include either one of the power switch 402 and thetimer 504. However, when the control module 400 includes the powerswitch 402, the second input unit 404 is also included; and when thecontrol module 400 includes the second input unit 404, the second outputunit 505 is also included.

Furthermore, the cancer treatment apparatus 100 may be configured sothat the control module 400 is not included and the magnetic fieldgenerator 200 generates the magnetic field at a fixed frequency of anyvalue from 100 kHz to 300 kHz. The configuration of the cancer treatmentapparatus 100 in this case is as illustrated in FIG. 28. Furthermore,the frequency of the magnetic field generated by the magnetic fieldgenerator 200 may be from 200 kHz to 300 kHz.

Furthermore, the control module 400 may control the temperature of theaffected tissues to become lower than a cancer cell killing temperature.Specifically speaking, a temperature range of 40° C. to 43° C. is knownas the cancer cells killing temperature, so that the control module 400may control the temperature of the affected tissues to become lower than40° C. In this case, the control module 400 specifies conditions toapply the magnetic field and duration when the magnetic field may becontinuously radiated for each cancer type, by referring to a databaseprepared in advance; and radiation duration, that is, the duration setto the timer 504 is set to be less than the duration obtained from thedatabase.

EXAMPLES

The present invention will be explained by using an example in which acopper-made, helical-type coil (inner diameter of a helical part: 4 cm;outer diameter of the helical part: 5 cm; the number of elements430,998/the number of nodes 137,055; and conductivity: 1.673e-8 [ohm m])is used as the magnetic field generator. Unless particularly explained,the electric current supplied to the magnetic field generator is 250 A.This example is Example 1. However, the present invention is not limitedto Example 1 as long as it exerts its operational advantages. No heatgeneration medium was used in any of the examples including Example 1.In other words, in none of the examples described below, no heatgeneration medium is placed around or inside the cells.

1. Magnetic Flux Density of Example 1

FIG. 6 is a simulation result of magnetic flux density of the magneticfield generator of Example 1. FIG. 6(b) shows the magnetic flux densitywhen the torso of the individual (mouse) was placed opposite ahelical-part inner diameter face 601 of a coil 600 of the magnetic fieldgenerator in a state illustrated in FIG. 6(a) and an electric current of335.4 A with the frequency of 280 kHz was applied. The distanceindicated by x-axis in FIG. 6(b) is a vertical distance from thehelical-part inner diameter face 601 of the coil 600 illustrated in FIG.6(a), that is, a 0-mm surface to the helical shaft center side. Five (5)mm along x-axis in FIG. 6(b) means an area 5 mm away from the 0-mmsurface towards the helical shaft center side (inside) as illustrated inFIG. 6(a). As a result of simulation of the magnetic flux density undermeasurement conditions for the magnetic flux density as described below,it was found that the increase of the distance as illustrated in FIG.6(b) caused a mild influence on the reduction of the magnetic fluxdensity and the magnetic flux density of the relevant area was 21 mT.When the magnetic field generator according to the present invention isplaced opposite the affected tissues in a non-contact state, thedistance between the magnetic field generator and the affected tissuesdoes not necessarily have to be strictly managed; and if it werenecessary to secure the magnetic flux density of 21 mT or more, it wouldbe preferable that the magnetic field generator and the affected tissuesbe placed opposite each other to apply the magnetic field to theaffected tissues in an area of 5 mm or less from the inner diameter faceof the coil.

<Measurement Conditions for Magnetic Flux Density>

Simulation Software: JMAG Designer 14.1

Solver: 3D FEM (Finite Element Method)

Simulation Type: Magnetic Field Analysis (Frequency Response)

2. Confirmation of Cancer Cell Cytostatic Effect

The cancer cell cytostatic effect of Example 1 was observed with respectto normal cells and cancer cells. The application was performed byplacing a petri dish, in which the relevant cells were sowed, inside thehelical part of the coil.

[2.1 Normal Cells]

FIG. 7 shows observation results of human stellate cells (NHA cells) asnormal cells to which Example 1 was applied. Cells purchased from Lonzagroup, Ltd. were used as the human stellate cells. FIG. 7(a) is an imagecaptured when cell culture was performed for 24 hours without applyingthe alternating magnetic field (Comparative Example 1-07a). FIG. 7(b) isan image captured when the cell culture was performed for 24 hours afterapplying the alternating magnetic field at the frequency of 196 kHz for30 minutes (Example 1-07b). FIG. 7(c) to FIG. 7(e) (Example 1-07c toExample 1-07e) are images captured by changing only the frequency in thecondition for FIG. 7(b) to 227 kHz, 280 kHz, and 389 kHz, respectively.In each of Example 1-07b to Example 1-07e and Comparative Example 1-07a,Culture Condition 1 described below was used to culture the cells. Theimages of FIG. 7(a) to FIG. 7(e) were captured by a fluorescencemicroscope NIKON TE2000E.

<Culture Condition 1>

Dyed 1×10⁵ cells/4-cm dish using Calcein-AM by Sigma (to dye livingcells) and propidium iodide (to dye nuclei and check dead cells) weresowed and cultured for 24 hours.

FIG. 7(f) is a graph showing an analysis result of the number of deadcells after the cell culture. The number of dead cells is the countnumber of cells by the fluorescence microscope. Five pieces of dataindicated in FIG. 7F correspond to FIG. 7(a) to FIG. 7(e), respectively.Specifically, the first “CTRL” is data based on the cell culture(Comparative Example 1-07a) explained about FIG. 7(a); the second “196kHz” is data based on the cell culture (Example 1-07b) explained aboutFIG. 7(b); the third “227 kHz” is data based on the cell culture(Example 1-07c) explained about FIG. 7(c); the fourth “280 kHz” is databased on the cell culture (Example 1-07d) explained about FIG. 7(d); andthe fifth “389 kHz” is data based on the cell culture (Example 1-07e)explained about FIG. 7(e).

No significant cytostatic effect could not be confirmed as illustratedin FIG. 7 even by using Example 1 on the human stellate cells (thenormal cells). Consequently, this result successfully confirmed that theeffect of suppressing the proliferation of the normal cells by thepresent invention was very low as compared to the cancer cell cytostaticeffect.

[2.2 Cancer Cells]

Experiments to check the cancer cell cytostatic effect of the presentinvention of Example 1 regarding the cancer cells were conducted abouthuman glioblastoma cells (U87), human malignant melanoma cells(SK-MEL-24), human tongue cancer (squamous cell neoplasm) cells(OSC-19), human breast cancer cells (MCF7), and human epithelial cellline derived from a lung carcinoma tissue (A549). An alternatingmagnetic field transistor inverter (Hot Shot, Ameritherm Inc., New York,U.S.A.) was used in the confirmatory experiments. The coil was asdescribed earlier.

[2.2.1 Human Glioblastoma Cells (U87)]

FIG. 8 shows observation results of the cytostatic effect by Example 1with respect to the human glioblastoma cells (U87). Human GB cell lineU87 cells purchased from American Type Culture Collection (ATCC)(Virginia, U.S.A.) were used as the human glioblastoma cells (U87). FIG.8(a) is an image captured when the cell culture was performed withoutapplying the alternating magnetic field (Comparative Example 1-08a).FIG. 8(b) is an image captured when the cell culture was performed afterapplying the alternating magnetic field at the frequency of 196 kHz for30 minutes (Example 1-08b). FIG. 8(c) to FIG. 8(e) (Example 1-08c toExample 1-08e) are images captured by changing only the frequency in thecondition for FIG. 8(b) to 227 kHz, 280 kHz, and 389 kHz, respectively.Culture Condition 1 was used to culture the cells and the images of FIG.8(a) to FIG. 8(e) were captured by the fluorescence microscope NIKONTE2000E.

FIG. 8(f) is a graph showing an analysis result of the number of deadcells after the cell culture. Five pieces of data indicated in FIG. 8Fcorrespond to FIG. 8(a) to FIG. 8(e), respectively. Specifically, thefirst “CTRL” is data based on the cell culture (Comparative Example1-08a) explained about FIG. 8(a); the second “196 kHz” is data based onthe cell culture (Example 1-08b) explained about FIG. 8(b); the third“227 kHz” is data based on the cell culture (Example 1-08c) explainedabout FIG. 8(c); the fourth “280 kHz” is data based on the cell culture(Example 1-08d) explained about FIG. 8(d); and the fifth “389 kHz” isdata based on the cell culture (Example 1-08e) explained about FIG.8(e). The number of dead cells was measured in the same manner as in theaforementioned case of the human stellate cells.

Regarding the human glioblastoma cells (U87), a significant increase ofthe number of dead cells was observed as illustrated in FIG. 8 byapplying the magnetic field at the frequencies of 196 kHz (Example1-08b), 227 kHz (Example 1-08c), and 280 kHz (Example 1-08d) usingExample 1. Consequently, this result confirmed that the presentinvention had the effect of suppressing the proliferation of the humanglioblastoma cells (U87).

[2.2.2 Human Malignant Melanoma Cells (SK-MEL-24)]

FIG. 9 shows observation results of the cytostatic effect by Example 1with respect to human malignant melanoma cells (SK-MEL-24). The humanmalignant melanoma cells (SK-MEL-24) purchased from ATCC were used. FIG.9(a) is an image captured when the cell culture was performed withoutapplying the alternating magnetic field (Comparative Example 1-09a).FIG. 9(b) to FIG. 9(e) are images captured respectively after applyingthe alternating magnetic field at the frequency of 196 kHz (Example1-09b) for FIG. 9(b), at the frequency of 227 kHz (Example 1-09c) forFIG. 9(c), at the frequency of 280 kHz (Example 1-09d) for FIG. 9(d),and at the frequency of 389 kHz (Example 1-09e) for FIG. 9(e) for 30minutes each time and then performing the cell culture. CultureCondition 1 was used to culture the cells and the images of FIG. 9(a) toFIG. 9(e) were captured by the fluorescence microscope NIKON TE2000E.

Five pieces of data indicated in FIG. 9(f) represent a graph indicatingthe analysis result of the number of dead cells after the culturecorresponding to FIG. 9(a) to FIG. 9(e) (Comparative Example 1-09a andExample 1-09b to Example 1-09e), respectively. The number of dead cellswas measured in the same manner as in the aforementioned case of thehuman stellate cells.

Regarding the human malignant melanoma cells (SK-MEL-24), a significantincrease of the number of dead cells was observed as illustrated in FIG.9 when the magnetic field was applied at the frequencies of 196 kHz(Example 1-09c), 227 kHz (Example 1-09d), and 280 kHz (Example 1-09e)using Example 1. This result confirmed that the present invention hadthe effect of suppressing the proliferation of the human malignantmelanoma cells (SK-MEL-24).

[2.2.3 Human Tongue Cancer (Squamous Cell Carcinoma) Cells (OSC-19)]

FIG. 10 shows observation results of the cytostatic effect by Example 1with respect to human tongue cancer (squamous cell carcinoma) cells(OSC-19). The human tongue cancer cells (OSC-19) purchased from TheHealth Science Research Resources Bank (Japan Health SciencesFoundation, Tokyo, Japan) were used. FIG. 10(a) is an image capturedwhen the cell culture was performed without applying the alternatingmagnetic field (Comparative Example 1-10a). FIG. 10(b) to FIG. 10(e) areimages captured respectively after applying the alternating magneticfield at the frequency of 196 kHz (Example 1-10b) for FIG. 10(b), at thefrequency of 227 kHz (Example 1-10c) for FIG. 10(c), at the frequency of280 kHz (Example 1-10d) for FIG. 10(d), and at the frequency of 389 kHz(Example 1-10e) for FIG. 10(e) for 30 minutes each time and thenperforming the cell culture. Culture Condition 1 was used to culture thecells and FIG. 10(a) to FIG. 10(e) were captured by the fluorescencemicroscope NIKON TE2000E.

Five pieces of data indicated in FIG. 10(e) represent a graph indicatingthe analysis result of the number of dead cells after the culturecorresponding to FIG. 10(a) to FIG. 10(e) (Comparative Example 1-10a andExample 1-10b to Example 1-10e), respectively. The number of dead cellswas measured in the same manner as in the aforementioned case of thehuman stellate cells.

Regarding the human tongue cancer cells (OSC-19) a significant increaseof the number of dead cells was observed as illustrated in FIG. 10 whenthe magnetic field was applied at the frequencies of 196 kHz (Example1-10b) and 227 kHz (Example 1-10c) using Example 1. This resultconfirmed that the present invention had the effect of suppressing thehuman tongue cancer cells (OSC-19).

[2.2.4 Human Breast Cancer Cells (MCF7)]

FIG. 11 shows observation results of the cytostatic effect by Example 1with respect to human breast cancer cells (MCF7). The human breastcancer cells (MCF7) purchased from ATCC were used. FIG. 11(a) is animage captured when the cell culture was performed without applying thealternating magnetic field (Comparative Example 1-11b). FIG. 11(b) toFIG. 11(e) are images captured respectively after applying thealternating magnetic field at the frequency of 196 kHz (Example 1-11b)for FIG. 11(b), at the frequency of 227 kHz (Example 1-11c) for FIG.11(c), at the frequency of 280 kHz (Example 1-11d) for FIG. 11(d), andat the frequency of 389 kHz (Example 1-11e) for FIG. 11(e) for 30minutes each time and then performing the cell culture. CultureCondition 1 was used to culture the cells and the images of FIG. 11(a)to FIG. 11(e) were captured by the fluorescence microscope NIKONTE2000E.

Five pieces of data indicated in FIG. 11(f) represent a graph indicatingthe analysis result of the number of dead cells after the culturecorresponding to FIG. 11(a) to FIG. 11(e) (Comparative Example 1-11a andExample 1-11b to Example 1-11e). The number of dead cells was measuredin the same manner as in the aforementioned case of the human stellatecells.

Regarding the human breast cancer cells (MCF7), a significant increaseof the number of dead cells was observed as illustrated in FIG. 11 whenthe magnetic field was applied at the frequencies of 196 kHz (Example1-11c) and 227 kHz (Example 1-11d) using Example 1. This resultconfirmed that the present invention had the effect of suppressing theproliferation of the breast cancer cells (MCF7).

[2.2.5 Human Epithelial Cell Line Derived from a Lung Carcinoma Tissue(A549)]

FIG. 20 shows observation results of the cytostatic effect by Example 1with respect to human epithelial cell line derived from a lung carcinomatissue (A549). The human epithelial cell line derived from a lungcarcinoma tissue (A549) purchased from ATCC were used. FIG. 20(a) is animage captured when the cell culture was performed without applying thealternating magnetic field (Comparative Example 1-20a). FIG. 20(b) toFIG. 20(e) are images captured respectively after applying thealternating magnetic field at the frequency of 196 kHz (Example 1-20b)for FIG. 20(b), at the frequency of 227 kHz (Example 1-20c) for FIG.20(c), at the frequency of 280 kHz (Example 1-20d) for FIG. 20(d), andat the frequency of 389 kHz (Example 1-20e) for FIG. 20(e) for 30minutes each time and then performing the cell culture. CultureCondition 1 was used to culture the cells and the images of FIG. 20(a)to FIG. 20(e) were captured by the fluorescence microscope NIKONTE2000E.

Five pieces of data indicated in FIG. 20(a) show a graph indicating theanalysis result of the number of dead cells after the culturecorresponding to FIG. 20(a) to FIG. 20(e) (Comparative Example 1-20a andExample 1-20b to Example 1-20e), respectively. The number of dead cellswas measured in the same manner as in the aforementioned case of thehuman stellate cells.

Regarding the human epithelial cell line derived from a lung carcinomatissue (A549), a significant increase of the number of dead cells wasobserved as illustrated in FIG. 20 when the magnetic field was appliedat the frequencies of 196 kHz (Example 1-20b) and 389 kHz (Example1-20e) using Example 1. This result confirmed that the present inventionhad the effect of suppressing the proliferation of the human epithelialcell line derived from a lung carcinoma tissue (A549).

The proliferation of the cancer cells is suppressed by applying themagnetic field at a specific frequency corresponding to the cancer type.Particularly, the present invention which has been illustrated by theexamples above can exert the excellent cancer cell cytostatic effectthrough a combination of the specific frequency and the applicationduration. Accordingly, not only the cancer cell cytostatic effect isenhanced, but also the reduction of the user's burden is realized bysetting the cancer type parameter, which is the combination of thespecific frequency and the application duration, and thereby making itpossible to easily apply a specified magnetic field. Therefore, thepresent invention should preferably be configured to store such aparameter, which is the combination of the specific frequency and theapplication duration, in the memory by associating the parameter withthe cancer type. Consequently, an input to the cancer type input unitevokes an arbitrary cancer type parameter, which is set as a desiredcancer type parameter, and the electric current is supplied to themagnetic field generator based on a set value. As a result, the presentinvention can generate an appropriate magnetic field to achieve thecytostatic effect for the desired cancer type and can easily achieve thecancer cell cytostatic effect by applying the magnetic field.

[2.3 Influence of Magnetic Field Application on Cell Division of CancerCells]

FIG. 12 shows observation results of a cell division state of the humanglioblastoma cells (U87). FIG. 12(a) is an image captured when the humanglioma cells (U87) were cultured without applying the magnetic field(Comparative Example 1-12a). FIG. 12(b) is an image captured afterapplying the alternating magnetic field at the frequency of 227 kHz for30 minutes by using Example 1 and then culturing the cells (Example1-12b). Culture Condition 1 was used to culture the cells and the imageswere captured by using the fluorescence microscope.

When the cells were cultured without applying the magnetic field(Comparative Example 1-12a), the cell division was observed at positionsindicated with white arrows as shown in FIG. 12(a). On the other hand,when the cells were cultured after applying the magnetic field (Example1-12b) as in FIG. 12(b), it was successfully observed that no celldivision occurred. Based on this observation result, it was possible toinfer that the cell division of the human glioblastoma cells (U87) wouldbe inhibited by using the present invention.

[2.4 Influence of Magnetic Field Application on Cancer Cells]

FIG. 13 shows scanning electron micrographs of the human glioblastomacells (U251).

FIG. 13(a) is an image captured when the human glioblastoma cells (U251)were cultured without applying the magnetic field by using Example 1(Comparative Example 1-13a). FIG. 13(b) is an image captured when thecells were cultured under Culture Condition 1 after applying themagnetic field at the frequency of 227 kHz for 30 minutes by usingExample 1 (Example 1-13b). As a result of comparison between FIG. 13(a)and FIG. 13(b), it was successfully observed as indicated with whitearrows in FIG. 13(b) that holes are made in the cell surfaces of Example1-1301. Based on this observation result, it was possible to infer thatcell surface membranes of the human glioblastoma cells (U251) could bedamaged by using the present invention.

[2.5 Temperature Change of Cancer Cells by Magnetic Field Application]

A subcutaneous tumor model was prepared by transplanting the humanglioblastoma cells (U87) at 5×10⁶/mouse under the skin of a femoralregion of a female 5-week-old Balb-c nude mouse. A transplanted area ofthe tumor model was placed opposite the magnetic field generator ofExample 1 and the magnetic field was applied to the transplanted area,and changes in the temperature of the tumor model were observed. Theapplication of the magnetic field was performed at the frequency of 227kHz for 30 minutes.

FIG. 14 shows observation results of the above-described tumor model.Referring to FIG. 14, an area with stronger whiteness has a highertemperature, so that changes in the temperature of the tumor model ascaused by the application of the magnetic field by using Example 1 canbe observed. The temperature was judged from image capture results bymeans of thermography. FIG. 14(a) is a temperature measurement result ofthe tumor model before the application. Regarding FIG. 14(a), thetemperature in the image was from 24° C. to 37.5° C. FIG. 14(b) is atemperature measurement result of the tumor model after the application.The highest temperature in the image was 36.1° C. and the temperature ofthe affected tissues surrounded by a broken line was 34.5° C.Specifically speaking, even when the magnetic field is applied by usingthe present invention, the temperature of the cancer cells does notreach the temperature range of 40° C. to 43° C., which is well-known asthe cancer cell killing temperature. Therefore, it was possible to inferthat the possibility for the cancer cell cytostatic effect according tothe present invention to be caused by killing of the cancer cells withhot heat would be low.

[2.6 Confirmation of Tumor Proliferation Suppressing Effect]

Tumor models of the human tongue cancer (squamous cell carcinoma) cells(OSC-19), the human glioblastoma cells (U87), the human breast cancer(MDAMB231), the human pancreatic cancer cells (PANC1), and the human themalignant mesothelioma cells (Meso-1) were prepared respectively and thetumor proliferation suppressing effect by the application of themagnetic field using Example 1 was checked. Enlargement of the tumor wasevaluated based on a volume change. Regarding the volume of the tumor,the tumor size of each individual of the tumor model was measured with avernier caliper and a value obtained by converting the tumor size intothe volume according to Expression (1) and Expression (2) was defined asthe tumor volume.

Tumor volume (TV) (mm³)=length×(width)²/2  (1)

Rate of tumor volume (%)=TV/TV_((day1))×100  (2)

[2.6.1 Tumor of Human Tongue Cancer (Squamous Cell Carcinoma) Cells(OSC-19)]

The human tongue cancer (squamous cell carcinoma) cells (OSC-19) weretransplanted at 1×10⁶/mouse under the skin of a femoral region of afemale 5-week-old Balb-c nude mouse in order to prepare subcutaneoustumor models using immunodeficient mice. The population was divided intotwo groups and the magnetic field was applied to individuals belongingto a first group by using Example 1. The first group was defined as anAMF group (Example 1-1501). The alternating magnetic field was appliedto the tumor of the AMF group (Example 1-1501) at the frequency of 227kHz for 30 minutes per day for consecutive five days from the 1st day tothe 5th day by setting the next day of the transplantation as the 1stday. No application was performed for individuals belonging to thesecond group. The second group was defined as a Control group(Comparative Example 1-1500). The tumor size of each individualbelonging to the AMF group (Example 1-1501) and the Control group(Comparative Example 1-1500) was measured every day and was convertedinto the tumor volume according to Expression (1) and Expression (2).

FIG. 15 shows observation results of changes in the tumor volume of thehuman tongue cancer (squamous cell carcinoma) cells (OSC-19).Exponentiation was performed by setting the tumor volume of the AMFgroup (Example 1-1501) and the Control group (Comparative Example1-1500) on the 1st day as 100 and FIG. 15(a) shows changes in the volumefor 15 days from the next day of the transplantation (the 1st day) withrespect to AMF (Example 1-1501) and Control (Comparative Example 1-1500)by using exponentiated volume values. As indicated in FIG. 15(a), theAMF group (Example 1-1501) was confirmed to have a tendency thatenlargement of their tumor volume on the 10th day was suppressed ascompared to the Control group (Comparative Example 1-1500). Anenlargement rate of the tumor of Control (Comparative Example 1-1500) onthe 10th day was 381%. On the other hand, the enlargement rate of thetumor of AMF (Example 1-1501) was 281%. As indicated in FIG. 15(a), theenlargement rate of the tumor of Control (Comparative Example 1-1500) onthe 15th day was 741%. On the other hand, the enlargement rate of thetumor of AMF (Example 1-1501) was 411%.

Referring to FIG. 15(b), the volume on the 14th day was exponentiated bydefining the volume on the 1st day as indicated in FIG. 15(a) as 100%and the enlargement of the tumor volume of Control (Comparative Example1-1500) was compared with that of AMF (Example 1-1501). As indicated inFIG. 15(b), the enlargement rate of the tumor of Control (ComparativeExample 1-1500) was 770%. On the other hand, the enlargement rate of thetumor of AMF (Example 1-1501) was 400%. This result successfullyconfirmed that the effect of suppressing the tumor proliferation of thetongue cancer (squamous cell carcinoma) continued for at least 14 daysby using the present invention.

[2.6.2 Tumor of Human Glioblastoma Cells (U87)]

FIG. 16 shows observation results of changes in the tumor volume of thehuman glioblastoma cells (U87). The human glioblastoma cells (U87)purchased from ATCC were used. Subcutaneous tumor models were preparedby transplanting the human glioblastoma cells U87 at 5×10⁶/mouse underthe skin of a femoral region of each female 5-week-old Balb-c nudemouse. The population was divided into three groups and the magneticfield was applied to individuals belonging to a first group once on thetransplantation day (the 0th day) by using Example 1. The first groupwas decided as Example 1-1601. The magnetic field was applied toindividuals belonging to a second group for consecutive 5 days from the0th day by using Example 1. The second group was decided as Example1-1605. The magnetic field was applied once per day for Example 1-1601and Example 1-1605 and in either case the frequency was 227 kHz and theapplication duration was 30 minutes. The magnetic field was not appliedto individuals belonging to a third group. The third group was decidedas Comparative Example 1-1600. The tumor size of each individualbelonging to Example 1-1601, Example 1-1605, and Comparative Example1-1600 was measured every day for consecutive 14 days from thetransplantation day (the 0th day) with the vernier caliper and wasconverted into the volume according to Expression (1) and Expression(2).

FIG. 16(a) is a captured image of individuals belonging to Example1-1601, Example 1-1605, and Comparative Example 1-1600 on the 14th day.Referring to FIG. 16(a), the inside of an area surrounded by a solidline is an affected part of each of the examples and the comparativeexample. Regarding the tumor size indicated within the solid line inFIG. 16(a), Comparative Example 1-1600 was the largest, Example 1-1601was smaller than Comparative Example 1-1600, and Example 1-1605 was thesmallest.

FIG. 16(b) is a graph indicating changes in the tumor volume of Example1-1601, Example 1-1605, and Comparative Example 1-1600 throughexponentiation by setting the volume on the 0th day as 100%. Asindicated in FIG. 16(b), the tumor enlargement rate of ComparativeExample 1-1600 after the 0th day relative to the volume on the 0th dayis 100% or more; and the volume enlargement tendency clearly appeared asdays elapsed. On the other hand, the tumor enlargement rate of Example1-1601 relative to the volume on the 0th day changed to 84.8% (the 2ndday), 105% (the 4th day), and 89.75% (the 7th day). Furthermore, theenlargement rate of Example 1-1605 relative to the volume on the 0th daychanged to 84.55% (the 2nd day), 89.44% (the 4th day), and 66.42% (the7th day). The enlargement tendency like Comparative Example 1-1600 wasnot observed with regard to the tumor volume of Example 1-1601 andExample 1-1605.

As a result of comparison of the changes in the tumor volume of eachexample and the comparative example indicated in FIG. 16(b), it wassuccessfully confirmed that the present invention had the effect ofsuppressing the proliferation of the glioblastoma tumor volume.Furthermore, as a result of comparison between Example 1-1601 andExample 1-1605, the enlargement rate of Example 1-1605 was lower onevery measurement day. These results successfully confirmed that whenthe magnetic field was applied once according to the present inventionunder the same application condition, the effect of suppressing theproliferation of the glioblastoma tumor volume was exerted better with alarger number of times of applications.

FIG. 16(b) shows measurement results from the 0th day to the 7th day,but the measurement also continued after the 7th day. FIG. 16(c) showsmeasurement results of the tumor volume of Example 1-1605 andComparative Example 1-1600 from the 0th day to the 14th day. The tumorvolume of Comparative Example 1-1600 was 152.8 mm³ on the 0th day, 206.8mm on the 2nd day, 295.8 mm³ on the 4th day, 329.6 mm³ on the 7th day,328.7 mm³ on the 9th day, 370.9 mm³ on the 11th day, and 752.8 mm³ onthe 14th day. On the other hand, the tumor volume of Example 1-1605 was181.5 mm³ on the 0th day, 153.3 mm³ on the 2nd day, 159.6 mm³ on the 4thday, 117.8 mm³ on the 7th day, 118.2 mm³ on the 9th day, 147.3 mm³ onthe 11th day, and 271.3 mm³ on the 14th day.

As indicated in FIG. 16(c), Example 1-1605 showed the significant cancercell cytostatic effect on and after the 4th day and its significantdifference from the volume of Comparative Example 1-1600 increased asdays elapsed. The changes in the tumor volume of Example 1-1605indicated in FIG. 16(c) successfully confirmed that the effect ofsuppressing the proliferation of glioblastoma tumor volume continued forat least 14 days by applying the magnetic field by using the presentinvention. It is also possible to infer that this effect would alsocontinue after the 14th day.

The present invention has the continuous effect of suppressing theproliferation of the tumor volume even with one application of themagnetic field. Also, the substantivity of the tumor volumeproliferation suppressing effect enhances as the number of times ofapplications is increased. As indicated in FIG. 16(b) and FIG. 16(c),when the magnetic field was applied once by using Example 1, the tumorvolume proliferation suppressing effect continued for at least 7 days;and when the magnetic field was applied 5 times, the tumor volumeproliferation suppressing effect continued for at least 14 days.

Therefore, the present invention may be configured so that the number oftimes of applications can be set by inputting the cancer type to thecancer type input unit. FIG. 17 shows an example of the cancer typeinput unit to which the parameter of the number of times of applicationscan be set. As illustrated in FIG. 17, the cancer type parameter isassociated with the frequency, the application duration, and the numberof times of applications which are stored in the memory; and specifiedfrequency, application duration, and the number of times of applicationscan be set and the magnetic field can be applied appropriately byinputting a desired cancer type. FIG. 17 illustrates an example of thecancer type input unit in which modes for the magnetic field applicationfor Example 1-1601 and Example 1-1605 can be set.

[2.6.3 Tumor of Human Breast Cancer (MDAMB231)]

A human breast cancer (MDAMB231) was transplanted at 5×10⁶/mouse underthe skin of a femoral region of a female 5-week-old Balb-c nude mouse inorder to prepare subcutaneous tumor models using immunodeficient mice.The population was divided into two groups and the magnetic field wasapplied to individuals belonging to a first group a total of 10 timesfor consecutive 5 days after the transplantation (from the 7th day), twodays with no application, and then again for consecutive 5 days by usingExample 1. The first group was decided as an AMF group (Example 1-2101).The magnetic field was applied once per day for Example 1-2101; andregarding each one application, the frequency was 227 kHz and theapplication duration was 30 minutes. The magnetic field was not appliedto individuals belonging to a second group. The second group was decidedas a CTRL group (Comparative Example 1-2100). The tumor size of eachindividual belonging to the AMF group (Example 1-2101) and the CTRLgroup (Comparative Example 1-2100) was measured every day forconsecutive 14 days after the transplantation (from the 7th day) withthe vernier caliper and was converted into the volume according toExpression (1) and Expression (2).

FIG. 21 is a diagram which shows the enlargement of the tumor volume bycomparing the CTRL group (Comparative Example 1-2100) with AMF (Example1-2101). Regarding the CTRL group (Comparative Example 1-2100), thevolume enlargement tendency clearly appeared as days elapsed. On theother hand, the volume of AMF (Example 1-2101) became smaller than, orstayed almost the same as, the 0th day. At any one of points in time,the tumor volume of AMF (Example 1-2101) was smaller than that of theCTRL group (Comparative Example 1-2100).

[2.6.4 Tumor of Human Pancreatic Cancer Cells (PANC1)]

Human pancreatic cancer cells (PANC1) were transplanted at 5×10⁶/mouseunder the skin of a femoral region of a female 5-week-old Balb-c nudemouse in order to prepare subcutaneous tumor models usingimmunodeficient mice. The population was divided into two groups and themagnetic field was applied to individuals belonging to a first group atotal of 10 times for consecutive 5 days after the transplantation (fromthe 7th day), two days with no application, and then again forconsecutive 5 days by using Example 1. The first group was decided as anAMF group (Example 1-2201). The magnetic field was applied once per dayfor Example 1-2201; and regarding each one application, the frequencywas 227 kHz and the application duration was 30 minutes. The magneticfield was not applied to individuals belonging to a second group. Thesecond group was decided as a CTRL group (Comparative Example 1-2200).The tumor size of each individual belonging to the AMF group (Example1-2201) and the CTRL group (Comparative Example 1-2200) was measuredevery day for consecutive 14 days after the transplantation (from the7th day) with the vernier caliper and was converted into the volumeaccording to Expression (1) and Expression (2).

FIG. 22 is a diagram which shows the enlargement of the tumor volume bycomparing the CTRL group (Comparative Example 1-2200) with AMF (Example1-2201). Regarding the CTRL group (Comparative Example 1-2200), thevolume enlargement tendency clearly appeared as days elapsed. On theother hand, the volume of AMF (Example 1-2201) continued decreasing asdays elapsed.

[2.6.5 Tumor of Human Glioblastoma Cells (U87) (Brain Tumor Models)]

FIG. 18 shows observation results of brain tumor models of the humanglioblastoma cells (U87). The operational advantages of the presentinvention were confirmed with fluorescent imaging and an overallsurvival rate of the tumor models as indicated in FIG. 18. The humanglioblastoma cells (U87) purchased from ATCC were used. Fireflyluciferase genes were transferred to the human glioblastoma cells byusing Lentiviruses by a known gene recombination method and then theobtained cells were cultured under Culture Condition 1, therebyproducing U87 cells transfected with a luciferase-encoding lentivirus.The brain tumor models were prepared by transplanting the U87 cellstransfected with a luciferase-encoding lentivirus at 1×10⁶/mouse to thebrain of a female 5-week-old Balb-c nude mouse. Subsequently,D-luciferin was administered to the brain tumor models to make itpossible to observe the tumor growth of U87 in the individuals. Theobservation terminated when the death of all the individuals wasconfirmed.

The population was divided into two groups and the magnetic field wasapplied to individuals belonging to a first group (the number ofindividuals: 6) by supplying an electric current of 250 A by usingExample 1. The first group was decided as Example 1-1805. The magneticfield was applied once per day for Example 1-1805; and regarding eachone application, the frequency was 227 kHz and the application durationwas 30 minutes. The magnetic field was applied for consecutive 5 daysfrom the 0th day (from the 0th day to the 4th day), no application wasapplied on the 5th day and the 6th day, and then the magnetic field wasapplied again from the 7th day to the 11th day. In other words, themagnetic field was applied to Example 1-1805 10 times. The magneticfield was not applied to individuals belonging to a second group (thenumber of individuals: 6). The second group was decided as ComparativeExample 1-1800.

Fluorescent imaging of Example 1-1805 and Comparative Example 1-1800 wasperformed by using an ultrasensitive CCD camera and an image analysisequipment (equipment name: IVIS imaging system). The tumor proliferationsuppressing effect was evaluated based on a value of the amount ofluminescence (photons/second); and a smaller amount of luminescence wasjudged to have the tumor proliferation suppressing effect. Onefluorescent image of the individuals belonging to Example 1-1805 and onefluorescent image of the individuals belonging to Comparative Example1-1800 were extracted respectively and are shown in FIG. 18(a). FIG.18(b) is a graph indicating changes in the amount of luminescence ofExample 1-1805 and Comparative Example 1-1800. As indicated in FIG.18(a) and FIG. 18(b), the amount of luminescence of Comparative Example1-1800 increased as days elapsed. On the other hand, the amount ofluminescence of Example 1-1805 did not show almost any increase evenwhen days elapsed; and particularly on and after the 16th day, theamount of luminescence was little as compared to Comparative Example1-1800 and a significant difference from Comparative Example 1-1800 wasobserved. Specifically speaking, it was successfully confirmed thatExample 1 had the effect of suppressing the proliferation of theglioblastoma tumor also with respect to the brain tumor models.

Furthermore, FIG. 18(c) shows an overall survival rate of Example 1-1805and Comparative Example 1-1800, which was calculated according toExpression (3).

Overall Survival Rate (%)=The Number of All Dead Individuals/6×100  (3)

Regarding Comparative Example 1-1800 as indicated in FIG. 18(c), thefirst individual died on the 20th day and all the individuals diedbefore and on the 28th day. The overall survival rate changed asfollows: 83% on the 20th day, 66% on the 22th day, 33% on the 25th day,16% on the 26th day, and 0% on the 28th day. On the other hand,regarding Example 1-1805, the first individual died on the 26th day andall the individuals died on the 35th day. The overall survival ratechanged as follows: 83% on the 26th day, 50% on the 27th day, 33% on the28th day, 16% on the 30th day, and 0% on the 35th day. It wassuccessfully confirmed that the application of a specified magneticfield by using Example 1 would provide an excellent prognosis of theglioblastoma.

Furthermore, human glioblastoma cells (U251) were used instead of thehuman glioblastoma cells (U87) and a similar observation was performed.Conditions other than the cell type are similar to those of the braintumor models of the human glioblastoma cells (U87) as indicated in FIG.18. The obtained overall survival rate is shown in FIG. 23. Regarding anuntreated group to which the magnetic field was not applied, the overallsurvival rate decreased on and after the 16th day; however, regarding analternating magnetic field group to which the magnetic field wasapplied, the overall survival rate of 100% was maintained even after 60days elapsed. Therefore, it was successfully confirmed that theapplication of a specified magnetic field by using Example 1 wouldprovide an excellent prognosis of the glioblastoma.

[2.6.6 Tumor of Malignant Mesothelioma Cells (Meso-1)]

FIG. 19 shows observation results of changes in the tumor volume ofhuman malignant mesothelioma cells (Meso-1). The human malignantmesothelioma cells (Meso-1) purchased from ATCC were used. Subcutaneoustumor models were prepared by transplanting the human malignantmesothelioma cells (Meso-1) at 1×10⁶/mouse under the skin of a buttockregion of each female 5-week-old Balb-c nude mouse. The population wasdivided into two groups and the magnetic field was applied toindividuals belonging to a first group on the transplantation day (the0th day) by using Example 1. The first group was decided as Example1-1901. The magnetic field was applied once for Example 1-1901 bysupplying the electric current of 250 A at the frequency of 308 kHzduring the application duration for 30 minutes. The magnetic field wasnot applied to individuals belonging to a second group. The second groupwas decided as Comparative Example 1-1900.

The tumor size of each individual belonging to Example 1-1901 andComparative Example 1-1900 was measured on the transplantation day, the2nd day, the 4th day, and the 6th day with the vernier caliper and wasconverted into the volume according to Expression (1) and Expression(2).

FIG. 19 is a graph indicating changes in the tumor volume of Example1-1901 and Comparative Example 1-1900 through exponentiation by settingthe volume on the 0th day as 100%. As indicated in FIG. 19, the tumorenlargement rate of Comparative Example 1-1900 after the 0th dayrelative to the volume on the 0th day is 100% or more; and the volumeenlarged as days elapsed. The enlargement rate of Comparative Example1-1900 relative to the volume on the 0th day changed as follows: 134%(the 2nd day), 163% (the 4th day), and 164% (the 6th day). On the otherhand, the enlargement rate of Example 1-1901 relative to the volume onthe 0th day changed as follows: 100% (the 2nd day), 107% (the 4th day),and 104% (the 6th day). The enlargement tendency like ComparativeExample 1-1900 was not observed with regard to the tumor volume ofExample 1-1901. Accordingly, it was successfully confirmed that thepresent invention had the effect of suppressing the tumor proliferationof the malignant mesothelioma.

3. Side Effects

Side effects caused by the applications of the magnetic field by usingthe magnetic field generator of Example 1 were observed. Female5-week-old Balb-c nude mice were divided into an AMF group to which themagnetic field was to be applied, and a CTRL group to which the magneticfield was to be not applied; and regarding each of the AMF group and theCTRL group, a group for transplanting a tumor under the skin and a groupfor transplanting the tumor inside the cranium were formed. The CTRLgroup will not be described below in detail, but correspondingindividuals were prepared for the comparison with the AMF group. The AMFgroup to which the tumor was transplanted under the skin was furtherdivided into a once-a-week group to which the magnetic field was appliedonce in a week, and a 5-times-a-week group to which the magnetic fieldwas applied five times in a week; and an observation was performed afterthe elapse of 15 days after the transplantation. Specifically speaking,regarding the once-a-week group of the AMF group with the subcutaneousimplant, the magnetic field was applied a total of twice; and regardingthe 5-times-a-week group of the same AMF group, the magnetic field wasapplied a total of 10 times. Regarding the AMF group with the tumortransplanted within the cranium, the magnetic field was applied fivetimes in a week, that is, a total of 40 times, and an observation wasperformed after the elapse of 90 days.

In any one of these cases, the magnetic field was applied for 30 minuteseach time and the frequency was 227 kHz. The observation was performedvia, for example, body weight measurement, blood drawing, and externalobservation; and specifically speaking, clinical signs, skin disorder,body weight, food intake, biological functions, kidney functions,panhemocytes, white blood cells, hemoglobin, blood platelets, and liverfunctions were evaluated.

FIG. 24 is a diagram indicating observation results of seven items, thatis, clinical signs, skin disorder, a reduction of body weight, areduction of food intake, degradation of biological functions,degradation of kidney functions, and a reduction of panhemocytes.However, regarding the reduction of the body weight, the reduction ofthe food intake, the degradation of the biological functions, thedegradation of the kidney functions, and the reduction of thepanhemocytes, whether a reduction or lowering of 20% or more from theCTRL group has occurred or not was observed. Regarding any one of theseitems, no side effects were observed as indicated in FIG. 24. Of theitems indicated in FIG. 24, the body weight will be described in detail.

FIG. 25 is a diagram showing changes in the body weight 7 days later and14 days later by setting the body weight on the observation start day as100%. Regarding each observation result of 7 days later and 14 dayslater, data of the CTRL group, the once-a-week group of the AMF group,and the 5-times-a-week group of the AMF group are indicated from theleft to the right. As indicated in FIG. 25, either one of the AMF groupshad a small difference in the body weight from the CTRL group; and noreduction of 20% or more from the CTRL group can be seen as indicated inFIG. 24.

FIG. 26 is a diagram showing results of the blood test after the elapseof 90 days with respect to the AF group with the tumor transplanted intheir cranium and the CTRL group. FIG. 26(a) to FIG. 26F sequentiallyshow white blood cells, hemoglobin, blood platelets, AST of the liverfunctions, and ALT of the liver functions, respectively. Regarding anyone of these test results, no significant difference between the CTRLgroup and the AMF group can be seen, so that no side effect can beacknowledged.

4. Reactive Oxygen

The influence of the application of the alternating magnetic field onreactive oxygen was observed with respect to the human glioblastomacells (U87), the human glioblastoma cells (U251), the human breastcancer cells (MDAMB231), and the human pancreatic cancer cells (PANC1).Each type of these cells was divided into an AMF group to which themagnetic field was applied, and a CTRL group to which the magnetic fieldwas not applied. Regarding the CTRL group, the alternating magneticfield was applied at 227 kHz for 30 minutes and the reactive oxygen wasmeasured after the elapse of 24 hours.

FIG. 29 is a diagram illustrating the influence of the application ofthe alternating magnetic field on the reactive oxygen with respect tothe human glioblastoma cells (U87), the human glioblastoma cells (U251),the human breast cancer cells (MDAMB231), and the human pancreaticcancer cells (PANC1). Regarding each one of these types of cells, ameasured value of the AMF group is indicated by setting a measured valueof the CTRL group as 100%. FIG. 29 shows that regarding each one of thehuman glioblastoma cells (U87), the human glioblastoma cells (U251), thehuman breast cancer cells (MDAMB231), and the human pancreatic cancercells (PANC1), the reactive oxygen increased significantly by applyingthe alternating magnetic field. For example, regarding the humanglioblastoma cells (U87), the reactive oxygen of the AMF group increasedtwice as much as that of the CTRL group.

The present invention exhibits the cancer cell cytostatic effect and thetumor proliferation suppressing effect even on different species fromthe species explained in the aforementioned examples, for example, onhumans. Specifically speaking, the present invention includes a methodfor treating human cancer cells by using the aforementioned cancertreatment apparatus 100. In other words, the present invention includesa human treatment method which is a method for treating humans by usingthe cancer treatment apparatus 100.

According to the present invention, the preferred alternating magneticfield for suppressing the tumor proliferation of various cancer typescan be applied to the affected tissues for preferred applicationduration by intuitive and simple operation of devices such as a keyboardand a mouse, and a touch panel. The application duration is short, andthe power consumption is small. In addition, the present invention islightweight. Accordingly, the present invention is a low-cost, highlyconvenient cancer treatment apparatus. Furthermore, since the affectedtissues does not contact the magnetic field generator, the cancertreatment apparatus according to the present invention can behygienically shared between a plurality of diseased persons.

INDUSTRIAL APPLICABILITY

The present invention is particularly suited for the remission treatmentof highly malignant cancers.

The disclosure of the following priority application is hereinincorporated by reference. Japanese Patent Application No. 2016-228164(filed on Nov. 24, 2016)

REFERENCE SIGNS LIST

-   100: cancer treatment apparatus-   200: magnetic field generator-   300: power source-   400: control module-   401: cancer type input unit-   402: power switch-   403: first input unit-   404: second input unit-   501: controller-   502: memory-   503: first output unit-   504: timer-   505: second output unit-   600: coil-   601: inner diameter face (0-mm surface) of coil

1. A cancer treatment apparatus for treating a cancer comprising: amagnetic field generator that generates a magnetic field of which afrequency is ranged from 100 kHz to 300 kHz to be applied to affectedtissues; a power source that generates an alternating current to besupplied to the magnetic field generator, and a control module thatcontrols the alternating current supplied from the power source to themagnetic field generator, wherein the control module controls thealternating current such that a magnetic flux density of the magneticfield, which is generated by the magnetic field generator, is apredetermined value, a frequency and size of the alternating currentsupplied to the magnetic field generator varies in correspondence withoutput from the control module,
 2. The cancer treatment apparatusaccording to claim 1, wherein the control module controls the powersource to maintain a temperature of the affected tissues lower than acancer cell killing temperature.
 3. The cancer treatment apparatusaccording to claim 1, wherein the affected tissues are human affectedtissues.
 4. The cancer treatment apparatus according to claim 1, whereinthe predetermined value of the magnetic flux density is at least 18 mTor larger. 5-10. (canceled)
 11. A treatment method for treating a cancercomprising: applying an alternating magnetic field to affected tissues.12. The treatment method for treating the cancer according to claim 11,wherein a frequency of the magnetic field is ranged from 100 kHz to 300kHz.
 13. The treatment method for treating the cancer according to claim11, wherein a frequency and size of the magnetic field is varied. 14.The treatment method for treating the cancer according to claim 11,further comprising without using any heat generation medium; and notusing heat-generating effect of the affected tissues due to theirradiation of the magnetic field for treatment.
 15. The treatmentmethod for treating the cancer according to claim 11, wherein a magneticflux density of the magnetic field is a predetermined.
 16. The treatmentmethod for treating the cancer according to claim 11, wherein thepredetermined value of the magnetic flux density is at least 18 mT orlarger.
 17. The treatment method for treating the cancer according toclaim 11, further comprising controlling a power source to generate analternating current to be supplied to a magnetic field generator suchthat the magnetic field is generated using the alternating current. 18.The treatment method for treating the cancer according to claim 11,further comprising: inputting a cancer type of the affected tissueswherein the cancer type, which is input, is defined as an input cancertype, determining the frequency of the magnetic field in correspondencewith the input cancer type.
 19. The treatment method for treating thecancer according to claim 11, wherein type of the cancer is aglioblastoma, a malignant melanoma, a tongue cancer, a breast cancer, amalignant mesothelioma, a pancreatic cancer, or a human alveolar basalepithelial adenocarcinoma.